Discovery of prolyl hydroxylase 2 inhibitors with new chemical scaffolds as in vivo active erythropoietin inducers through a combined virtual screening strategy

Abstract

Hypoxia-inducible factor (HIF) is identified to be a promising target to mediate the response to hypoxia. Its stability and activation are negatively controlled by prolyl hydroxylase 2 (PHD2). Thus, PHD2 inhibition has been perceived as a promising anti-anemia therapy. In this study, we carried out a structure-based virtual screening followed by in vitro and in vivo biological validation, with the goal to identify novel PHD2 inhibitors. As a result, a set of hits with new chemical scaffolds were revealed to be active in vitro for PHD2 inhibition. Compounds 2 and 3 were revealed to be capable of stabilizing HIF-α and stimulating erythropoietin (EPO) expression in cell-based assays. Notably, further in vivo assays revealed that 2 was capable of elevating the EPO plasma levels in C57BL/6 mice model. These findings provide new chemical scaffolds for further development of PHD2 inhibitors. K E Y W O R D Santi-anemia, EPO, HIF, PHD2 inhibitor, virtual screening SUPPORTING INFORMATIONAdditional supporting information may be found online in the Supporting Information section. How to cite this article: Yu Z, Li Z, Yu Q, et al. Discovery of prolyl hydroxylase 2 inhibitors with new chemical scaffolds as in vivo active erythropoietin inducers through a combined virtual screening strategy. Chem Biol Drug Des. 2020;95:270-278.

Dihydroazolopyrimidines: Past, Present and Perspectives in Synthesis, Green Chemistry and Drug Discovery

“…Dihydro‐1,2,4‐triazolo[1,5 ‐a ]pyrimidine 45 (Figure 13) was selected as a potential PHD2 inhibitor from the commercial compound database by a structure‐based virtual screening strategy combined with pharmacophore modeling and molecular docking. However, biological evaluations showed that the potency of compound 284 (IC 50 =12.79±0.38 μM) was lower than that of the approved drug roxadustat (IC 50 =0.59±0.02 μM) [334] …”

“…However, biological evaluations showed that the potency of compound 284 (IC 50 = 12.79 � 0.38 μM) was lower than that of the approved drug roxadustat (IC 50 = 0.59 � 0.02 μM). [334]…”

Fabricating Multiphasic Angiogenic Scaffolds Using Amyloid/Roxadustat-Assisted High-Temperature Protein Printing

“…In the latter study, although the concentration of 50 μM was found to be inhibitory, in another study, 57 this concentration was found to be suitable for stabilizing HIF-1α, therefore, for better angiogenesis. While these amounts of the drug were investigated on the cellular impact, in a study, 58 the inhibitory role of FG-4592 with different concentrations of prolyl hydroxylase 2 (PHD2) was directly investigated. It was seen that by increasing the concentration up to 150 μM, the inhibitory role also increases, but the generalization of this response to the cellular role of the drug cannot be correct.…”

Development of a colorimetric α-ketoglutarate detection assay for prolyl hydroxylase domain (PHD) proteins

“…As one application of this assay may be for a highthroughput screen for inhibitors of PHDs, we performed a proof-of-concept experiment by testing whether it is sensitive enough to detect known PHD2 inhibitors in ongoing clinical trials. At present, there are a number of small-molecule inhibitors of PHD2 in ongoing clinical trials to treat CKDinduced anemia (47). We selected three commercially available inhibitors in current phase 3 trials to evaluate the sensitivity of the 2,4-DNPH α-KG assay: Vadadustat (AKB-6548 from Akebia, approved in Japan), Roxadustat (FG-4592 from FibroGen, approved in Japan and China), and Daprodustat (GSK1278863 from GSK).…”